Human-controlled reproductive experience may contribute to incestuous behavior observed in reintroduced semi-feral stallions (Equuscaballus).

Equine reproductive behavior is affected by many factors, some remaining poorly understood. This study tested the hypothesis that a period of captivity during the juvenile period and human-controlled reproduction may potentially be involved in the disruption of the development of incestuous mating avoidance behavior in sanctuary-reintroduced male Konik polski horses. Between 1986 and 2000, cases of incestuous behavior in harem stallions born and reared until weaning in the sanctuary were studied. Eight males lived in the sanctuary's feral herd for the rest of their lives (the non-captive group; nC). They gained their own harem of mares without human intervention (no human-controlled reproductive activity, nHC). Another five stallions were removed as weanlings, reared in captivity and then reintroduced as adults (captive, C). Three of these C stallions were used as in-hand breeding stallions, one as a "teaser" (human-controlled reproductive activity, HC) and one was not used for reproduction in captivity (nHC). Reproductive records for 46 mares, daughters of all 13 harem stallions, were scrutinized and cases of incestuous breeding were recorded by interrogation of foal parentage records. C stallions failed to expel more daughters than nC stallions (33% vs. 18%, P = 0.045), and mated with significantly more of them (28% vs. 11%, P = 0.025). Interestingly, HC stallions expelled fewer (60%) and successfully mated with more (33%) daughters that nHC stallions (84% expelled, P = 0.013, and 10% successful mating with daughters, P = 0.010). All HC stallions bred incestuously at least once. We propose that human intervention during a critical period of development of social and reproductive behavior in young stallions, by enforced separation from their natal herd and in-hand breeding, may contribute to their later aberrant behavior and disruption of inbreeding avoidance mechanisms in these stallions. The previous occurrence of human-controlled breeding may be one of the factors promoting incestuous behavior of stallions in natural conditions. The uninterrupted presence of stallions in their harems and herd member recognition may also play important roles in inbreeding avoidance in horses.

Some Genetic and Environmental Effects on Equine Asthma in Polish Konik Horses.

Current knowledge of the genetic and environmental backgrounds of equine asthma seems to be insufficient, especially for primitive horse breeds. The main objectives of this study were to estimate the effects of sex, birth period, stud, parentage line and inbreeding on asthma morbidity in Polish Konik horses. Records of 274 horses (housed in two studs) were analysed. These animals were allocated to maternal and paternal lines. Individual inbreeding coefficients were extracted from the additive relationship matrix. Horses underwent diagnosis based on observation of the basic symptoms (high frequency of coughing and excessive nasal discharge). Subsequently, some horses (28 individuals) were clinically examined to confirm the earlier observations. Generally, no significant effects of parentage line on heaves morbidity were identified by the use of logistic regression, although the Pearson's chi-squared test had shown that individuals of some maternal and paternal lines had a predisposition to severe equine asthma. It was concluded that the individual inbreeding level is not associated with the incidence of EA, but a significant effect of the maternal inbreeding coefficient may be observed. It was also found that there is some variability in the incidence of this disease between studs.

The examination of biophysical parameters of the skin in Polish Konik horses.

This study aimed to assess the biophysical parameters of the skin in Polish Konik horses (Polish primitive horses). According to the authors, this is the first assessment performed on such a wide scale in this group of animals. The evaluation carried out is innovative both with regards to the breed of the animals and the wide scope of the physicochemical skin assessment. The study group comprised mares, stallions and geldings, and the evaluations concerned transepidermal water loss, corneometry, pH, skin temperature assessment and mexametry. These parameters were assessed in five skin regions: the lips, the right ear, the prosternum, the right side of the neck and the chest. The measurements were taken after spreading the hair apart, with the use of a Multiprobe Adapter System (MPA®) and dedicated probes (Courage + Khazaka electronic GmbH, Cologne, Germany). The measurements revealed statistically significant differences in the values of transepidermal water loss in the lips in mares compared with stallions (P = 0.023) and also in stallions compared with geldings (P = 0.009). Corneometry showed significantly higher results in the neck region in mares compared with stallions (P = 0.037) and the prosternum areas in mares and geldings compared with stallions (P = 0.037 and P = 0.018). Skin pH measurement on the right side of the neck rendered significantly higher values in stallions than in mares (P = 0.037). In geldings, the skin temperature was significantly higher than in stallions (P = 0.049). Once the appropriate physicochemical values for specific animal species and breeds are determined, non-invasive methods of skin examination in many diseases and also methods of evaluation of the efficacy and/or adverse effects of applied medications can be established.

Welfare of Free-Roaming Horses: 70 Years of Experience with Konik Polski Breeding in Poland.

To prevent abuse and to assure the welfare of domestic horses, attempts to assess welfare in a standardized way have been made. Welfare-assessment tools often refer to the physical and social environments of feral domestic horses as examples of welfare-friendly conditions for horses. However, free-roaming horses are often exposed to conditions or states that may be regarded as welfare threats or abuse. The aim of this review was to present cases of welfare compromises as well as natural ways to restore high standards of welfare to Konik polski horses (Koniks) living in semiferal conditions in a forest sanctuary over the course of 70 years. Welfare problems in Koniks related to feeding, locomotor, social, reproductive, and comfort behavior, as well as health issues concerning hoof trimming and parasitism in Koniks, are discussed. Periodic food scarcity or abundance, stressful events around weaning and gathering, the consequences of fights among stallions, exposure to sire aggression during dispersal, lameness during "self-trimming," exposure to insect harassment, high levels of parasitism, and specific landscape formations may endanger free-roaming horses. It has to be underlined that despite the excellent adaptability of horses to free-roaming conditions, one should be aware that welfare problems are to be expected in any semiferal population. Here, we present the management system applied for 70 years in free-roaming Konik polski horses that minimizes welfare threats. It allows close follow-up of individual horses, the strict monitoring of health and welfare on a daily basis, and if necessary, instant reactions from caretakers in cases of emergency. Moreover, it addresses the problem of starvation due to overgrazing and thus, the ethical controversy related to the eradication of surplus animals causing environmental damage.

Harem stallion changes are not associated with diminished reproductive performance of females in semi-feral Konik polski horses (Equus caballus).

Infanticide, related to a stallion's aggression toward a foal sired by another stallion, and feticide related to a new stallion's aggression and/or pheromonal influence (the Bruce effect) inducing loss of a fetus sired by another stallion, a female's counteraction to infanticide, have been proposed for domestic horses (Equus caballus) in human-managed conditions. The aim of the present study was, in conditions close to natural, to investigate the influence of the natural succession of a harem stallion on the mares' subsequent reproductive performance. In a population of semi-feral Konik polski horses observed for 31 years (reproductive seasons) in 8 bands, harem stallion changed 10 times. These changes involved 26 out of 48 mares and 60 out of 609 observed mare-seasons (MS, a year in which a mare experienced a reproductive event). Binary distribution and log link function were assumed. The marginal model included the classification variable (SCH) and the continuous variables (age of the mare and calendar year of reproductive event (birth of a live foal, abortion, foals lost or barrenness) in a given MS was analyzed with generalized linear mixed model. The reproductive fitness of mares and their reproductive success (foal surviving ≥ 1 year), did not differ between MS with and without SCH. Older females were more likely to stay barren, with chances increasing by 21% with each successive year; and less likely to give birth to a foal (13% decrease of chance), and rear a foal to one year of age (12% decrease of chance). The age did not affect the probability of abortions. Of the 26 MS when mares were pregnant when the stallion had changed, there were 25 healthy foals born. For the entire 31 years of monitoring, no aggression toward any foal was observed and all foals that were born in the harem of a new, succeeding stallion successfully reached adulthood. Due to the lack of incidents of infanticide and the lack of evidence suggesting that the presence of a new harem stallion leads to the termination of pregnancies sired by another stallion, the Bruce effect was not confirmed as a biological strategy to reduce investment in pregnancy and potential infanticide in studied population of semi-feral horses.

Do olfactory behaviour and marking responses of Konik polski stallions to faeces from conspecifics of either sex differ?

The aim of the study was to assess whether adult stallions differentiate their olfactory and marking behaviour towards the excreta of their potential male rivals and mares. Four Konik polski stallions were individually exposed simultaneously to their own and each others' faeces as well as faeces of mares in oestrus vs. dioestrus. Five series of 30min observation sessions were conducted in a round pen each on two consecutive days from March to July, totaling in 5h observation per stallion. Stallions sniffed and displayed flehmen reaction towards mares' faeces significantly longer (P<0.01) than towards stallions' faeces. No significant differences were found in marking by defecations upon stallions' vs. mares' faeces. The stallions urinated exclusively on mares' faeces. Sniffing, flehmen, defecations and urinations performed towards faeces of mares in oestrus vs. dioestrus did not differ significantly. The results showed that stallions differentiate their olfactory behaviour towards excreta of conspecifics of different sex but perhaps not towards faeces of females in different reproductive status. This for the first time establishes that stallions exhibit different marking behaviour upon stallions' vs. mares' faeces. It is hypothesized that marking of other stallions' faeces by defecation may advertise presence of the stallion, whereas marking of mares' faeces by urination may serve to mask the presence of a mare to prevent potential rivals from locating the mare in an area where mares' faeces are found.

Serum 8-hydroxy-2-deoxyguanosine as a marker of DNA oxidative damage in horses with recurrent airway obstruction.

BACKGROUND: It has been reported that equine recurrent airway obstruction (RAO) is a state of oxidative stress. Oxidant-antioxidant imbalance is known to increase the conversion of deoxyguanosine to 8-hydroxy-2-deoxyguanosine (8-OHdG) in DNA. 8-OHdG can easily be measured using ELISA tests in serum or urine samples. In this study, we analysed serum 8-OHdG levels in horses with recurrent airway obstruction and in healthy controls. RESULTS: The study material consisted of seven healthy horses and seven horses with symptomatic RAO. All horses were exposed to moldy hay and straw for 48 h to induce clinical exacerbation of RAO. The serum 8-OHdG levels were determined using the ELISA Highly Sensitive 8-OHdG kit. The difference between the levels of 8-OHdG in healthy and RAO-affected horses was significant. The median level of 8-OHdG was 0.044 ng/ml in the healthy controls versus 0.498 ng/ml in RAO horses (P = 0.0021). CONCLUSIONS: The results of the study strongly suggest that DNA damage coexists in the course of equine RAO. We therefore propose that future research should aim at the development of new drugs that target pro-inflammatory molecules, since DNA damage appears to be the result of chronic inflammation.

Serum concentrations of allergen-specific IgE in horses with equine recurrent airway obstruction and healthy controls assessed by ELISA.

BACKGROUND: Equine recurrent airway obstruction (RAO), also known as heaves, is one of the most common respiratory problems in older horses. When RAO-affected horses stay pastured or in a dust-free environment for a prolonged time, clinical signs as well as airway inflammation wane. A number of environmental, immunologic, infectious, and genetic factors play an important role in the pathogenesis of RAO, and the immunologic basis of this disease is still poorly understood. OBJECTIVES: The aim of this study was to investigate the concentrations of allergen-specific IgE in the serum of horses suffering from RAO and healthy controls. MATERIAL AND METHODS: The study included a group of 14 adult Polish Konik horses, kept in a standardized environment, and divided into 2 groups: 7 horses which did not have any respiratory problems comprised the control group and 7 horses with a history of RAO constituted the study group. A clinical and laboratory evaluation, endoscopic examination, and bronchoalveolar lavage (BAL) were performed in all horses. Sera of all horses were tested against allergens from 9 molds and 3 mites using the Heska Allercept assay. RESULTS: In the serologic tests, a statistically significant difference between both groups was found for specific IgE against mites, wherein Tyrophagus putrescentia correlated most clearly with RAO. There was no difference between groups for IgE specific against molds. CONCLUSION: On the basis of our observations and results, we conclude that RAO is associated with increased serum concentrations of specific serum IgE against mites, in particular T putrescentia.

Fuzzy logic-based diagnostic algorithm for implantable cardioverter defibrillators.

OBJECTIVE: The paper presents a diagnostic algorithm for classifying cardiac tachyarrhythmias for implantable cardioverter defibrillators (ICDs). The main aim was to develop an algorithm that could reduce the rate of occurrence of inappropriate therapies, which are often observed in existing ICDs. To achieve low energy consumption, which is a critical factor for implantable medical devices, very low computational complexity of the algorithm was crucial. The study describes and validates such an algorithm and estimates its clinical value. METHODOLOGY: The algorithm was based on the heart rate variability (HRV) analysis. The input data for our algorithm were: RR-interval (I), as extracted from raw intracardiac electrogram (EGM), and in addition two other features of HRV called here onset (ONS) and instability (INST). 6 diagnostic categories were considered: ventricular fibrillation (VF), ventricular tachycardia (VT), sinus tachycardia (ST), detection artifacts and irregularities (including extrasystoles) (DAI), atrial tachyarrhythmias (ATF) and no tachycardia (i.e. normal sinus rhythm) (NT). The initial set of fuzzy rules based on the distributions of I, ONS and INST in the 6 categories was optimized by means of a software tool for automatic rule assessment using simulated annealing. A training data set with 74 EGM recordings was used during optimization, and the algorithm was validated with a validation data set with 58 EGM recordings. Real life recordings stored in defibrillator memories were used. Additionally the algorithm was tested on 2 sets of recordings from the PhysioBank databases: MIT-BIH Arrhythmia Database and MIT-BIH Supraventricular Arrhythmia Database. A custom CMOS integrated circuit implementing the diagnostic algorithm was designed in order to estimate the power consumption. A dedicated Web site, which provides public online access to the algorithm, has been created and is available for testing it. RESULTS: The total number of events in our training and validation sets was 132. In total 57 shocks and 28 antitachycardia pacing (ATP) therapies were delivered by ICDs. 25 out of 57 shocks were unjustified: 7 for ST, 12 for DAI, 6 for ATF. Our fuzzy rule-based diagnostic algorithm correctly recognized all episodes of VF and VT, except for one case where VT was recognized as VF. In four cases short lasting, spontaneously ending VT episodes were not detected (in these cases no therapy was needed and they were not detected by ICDs either). In other words, a fuzzy logic algorithm driven ICD would deliver one unjustified shock and deliver correct therapies in all other cases. In the tests, no adjustments of our algorithm to individual patients were needed. The sensitivity and specificity calculated from the results were 100% and 98%, respectively. In 126 ECG recordings from PhysioBank (about 30min each) our algorithm incorrectly detected 4 episodes of VT, which should rather be classified as fast supraventricular tachycardias. The estimated power consumption of the dedicated integrated circuit implementing the algorithm was below 120nW. CONCLUSION: The paper presents a fuzzy logic-based control algorithm for ICD. Its main advantages are: simplicity and ability to decrease the rate of occurrence of inappropriate therapies. The algorithm can work in real time (i.e. update the diagnosis after every RR-interval) with very limited computational resources.

Neutrophil and macrophage apoptosis in bronchoalveolar lavage fluid from healthy horses and horses with recurrent airway obstruction (RAO).

BACKGROUND: Dysregulation of apoptosis has been implicated in a range of diseases including tumors, neurodegenerative and autoimmune diseases, as well as allergic asthma and chronic obstructive pulmonary disease (COPD) in humans. Although it has a different pathophysiology, delayed apoptosis of various inflammatory cells may play a pivotal role in the development of recurrent airway obstruction (RAO) in horses. Reduction of inflammatory cell apoptosis or a dysregulation of this process could lead to chronic inflammation and tissue injury. Therefore, the aim of this study was to investigate the rate of apoptosis and necrosis of neutrophils and macrophages in bronchoalveolar lavage fluid obtained from seven horses suffering from RAO (study group) and seven control horses. RESULTS: We demonstrated that neutrophil/macrophage apoptosis is altered in RAO-affected horses compared with the control group in the BAL fluid. We found a significant difference between the median percentage of early and late apoptosis of neutrophils between the study and control group of horses. Moreover, we found a positive correlation between the rate of apoptosis and the median percentage of macrophages in RAO-affected horses. CONCLUSION: The findings suggest that apoptosis dysregulation may play a significant role in the pathogenesis of RAO. However, further studies are needed to clarify the role of altered apoptosis in the course of equine recurrent airway obstruction.

Serum biochemical reference intervals for the Polish Konik horse (Equus cabalus gmelini Ant.).

BACKGROUND: Establishment of reference intervals (RI) for serum biochemical analytes is important for monitoring the health of different breeds of horses. OBJECTIVES: The aim of this study was to measure and report RI for serum biochemical analytes of the Polish Konik horse. MATERIAL AND METHODS: Blood samples were collected from 74 clinically healthy Polish Konik horses living under controlled natural conditions. These were adult primitive horses, aged 3-15 years, including 28 males (21 stallions, 7 geldings) and 46 mares. Serum analytes were measured and analyzed using a commercial automated analyzer. RESULTS: The following RI (medians) were comparable to previously published RI in horses: albumin 34.8-41.3 g/L (38.0); ALP 124-309 U/L (216); ALT 6-33 U/L (19); AST 300-566 U/L (433); calcium 2.8-3.2 mmol/L (3.0); chloride 95-102 mmol/L (99); cholesterol 2.1-3.4 mmol/L (2.7); CK 167-488 U/L (328); creatinine 101-170 µmol/L (136); glucose 2.7-5.3 mmol/L (4.0); lipase 303-1143 U/L (723); magnesium 0.8-1.0 mmol/L (0.9); phosphate 0.9-1.5 mmol/L (1.2); potassium 2.4-5.0 mmol/L (3.7); sodium 138-144 mmol/L (141); total bilirubin 8-16 µmol/L (11.7); total protein 63.5-78.9 g/L (71.2); triglycerides 0.0-0.3 mmol/L (0.1); urea 2.2-7.3 mmol/L (4.7). CONCLUSION: Reference intervals established in this study provide a valuable preliminary baseline for assessment of serum analytes in healthy Polish Konik horses.

Differences in seasonal changes of fecal androgen levels between stabled and free-ranging Polish Konik stallions.

Blood and feces samples were collected from Polish Konik stallions kept under conventional stable conditions and in the forest reserve during a 1-year study period. Levels of testosterone (T) and androstenedione (A(4)) were measured using radioimmunoassay. Positive correlation between fecal and plasma concentrations of androgens was observed. Fecal T concentrations increased in April and May reaching peak value mid-April in the stallions from the reserve group and 2 weeks later in the stallions from the stable group. Comparatively, concentrations of T were higher in the stable group. Levels of T decreased in July, increasing through August to September, and decreasing again in October. During this period of increase, concentrations of T were higher in the reserve group. A peak of fecal A(4) concentrations in the reserve group was noted mid-April, but high levels of this androgen appeared later and remained longer (also in May). When the breeding season finished, the concentration of A(4) decreased and remained low. During breeding season, concentrations of A(4) were higher in fecal samples taken from stable stallions than from those in the reserve group. During non-breeding seasons levels of this androgen in both groups were similar. The individual differences in the fecal androgen levels were dependent on the behavior of the stallions and were not related to their age. The seasonal changes of fecal T and A(4) levels differed between stabled and free-ranging Polish Konik stallions.

Serologic survey of potential vertebrate hosts for West Nile virus in Poland.

A survey for antibodies to West Nile virus (WNV; genus ,Flavivirus) was carried out by plaque-re-duction neutralization microtesting in 78 horses, 20 domestic chickens, and 97 wild birds belonging to 10 species from different areas in Poland. Specific antibodies were detected in five juvenile (hatching-year) birds collected in 2006: three white storks (Ciconia ciconia) in a wildlife rehabilitation center (5.4% of all examined storks; the antibody titers in each bird were 1:320, 1:160, and 1:20), one free-living mute swan (Cygnus olor; the titer was 1:20), and one hooded crow (Corvus corone cornix; the titer 1:20) in a wildlife rehabilitation center; thus the overall seropositivity to WNV was 5.2% among all the birds sampled. These data do not rule out the presence of WNV activity in Poland with 100% certainty, but they indicate a significant trace that demands verification. In addition, one black-headed gull (Larus ridibundus) had neutralizing antibodies for the Usutu Flavivirus, the first case recorded in Poland.